Regenerative Endodontics| Volume 47, ISSUE 6, P924-931, June 2021

Download started.


Vascular Endothelial Growth Factor and/or Nerve Growth Factor Treatment Induces Expression of Dentinogenic, Neuronal, and Healing Markers in Stem Cells of the Apical Papilla

Published:February 26, 2021DOI:



      The goal of regenerative endodontic procedures is to preserve and stimulate stem cells from the apical papilla (SCAPs) to develop the pulp-dentin complex using various growth factors and scaffolds. We hypothesized that the treatment of SCAPs with vascular endothelial growth factor (VEGF) or nerve growth factor (NGF) may impact the expression of osteogenic and dentinogenic markers.


      The optimum concentration of VEGF and NGF on SCAP viability was assessed and introduced to SCAPs for 6–24 hours. SCAPs were also challenged with Escherichia coli lipopolysaccharide (LPS). Messenger RNA (mRNA) expression of DSPP, DMP1, TGFB1, OCN, SP7, and TWIST1 was examined via quantitative reverse transcription polymerase chain reaction. Immunohistochemistry was used to verify protein expression. In addition, total RNA from NGF-treated SCAPs in the presence or absence of LPS was extracted for RNA sequencing.


      Compared with untreated cells, NGF-treated SCAPs showed markedly higher levels of DSPP, DMP1, and TGFB1 mRNAs (>9-fold change, P < .05), and SCAPs treated with both VEGF and NGF showed a significant increase of DSPP and TGFB1 mRNAs (P < .05). In addition, in LPS-challenged SCAPs, treatment with these growth factors also exhibited increased expression of DSPP, DMP1, and TGFB1 mRNAs, with the most significant change induced by VEGF (P < .05). Immunohistochemistry confirmed increased dentin sialophosphoprotein, dentin matrix acidic phosphoprotein 1, and transforming growth factor beta 1 protein expression in treated SCAPs. RNA sequencing revealed multiple pathways regulated by NGF, including TGF-β and neurogenic pathways.


      VEGF- and NGF-induced dentinogenic/neuronal/healing marker expression in SCAPs indicates the potential value of applying these growth factors in regenerative endodontic procedures.

      Key Words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Journal of Endodontics
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Diogenes A.
        • Ruparel N.B.
        Regenerative endodontic procedures: clinical outcomes.
        Dent Clin North Am. 2017; 61: 111-125
        • Sonoyama W.
        • Liu Y.
        • Yamaza T.
        • et al.
        Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study.
        J Endod. 2008; 34: 166-171
        • Sonoyama W.
        • Liu Y.
        • Fang D.
        • et al.
        Mesenchymal stem cell-mediated functional tooth regeneration in swine.
        PLoS One. 2006; 1: e79
        • Lei L.
        • Chen Y.
        • Zhou R.
        • et al.
        Histologic and immunohistochemical findings of a human immature permanent tooth with apical periodontitis after regenerative endodontic treatment.
        J Endod. 2015; 41: 1172-1179
        • Murray P.E.
        • Garcia-Godoy F.
        • Hargreaves K.M.
        Regenerative endodontics: a review of current status and a call for action.
        J Endod. 2007; 33: 377-390
        • Bottino M.C.
        • Pankajakshan D.
        • Nor J.E.
        Advanced scaffolds for dental pulp and periodontal regeneration.
        Dent Clin North Am. 2017; 61: 689-711
        • Steed D.L.
        The role of growth factors in wound healing.
        Surg Clin North Am. 1997; 77: 575-586
        • Folkman J.
        • Shing Y.
        J Biol Chem. 1992; 267: 10931-10934
        • Bussolino F.
        • Mantovani A.
        • Persico G.
        Molecular mechanisms of blood vessel formation.
        Trends Biochem Sci. 1997; 22: 251-256
        • Ucuzian A.A.
        • Gassman A.A.
        • East A.T.
        • Greisler H.P.
        Molecular mediators of angiogenesis.
        J Burn Care Res. 2010; 31: 158-175
        • Mitsiadis T.A.
        • Magloire H.
        • Pagella P.
        Nerve growth factor signalling in pathology and regeneration of human teeth.
        Sci Rep. 2017; 7: 1327
        • Mahdee A.
        • Eastham J.
        • Whitworth J.M.
        • Gillespie J.I.
        Evidence for changing nerve growth factor signalling mechanisms during development, maturation and ageing in the rat molar pulp.
        Int Endod J. 2019; 52: 211-222
        • Byers M.R.
        • Schatteman G.C.
        • Bothwell M.
        Multiple functions for NGF receptor in developing, aging and injured rat teeth are suggested by epithelial, mesenchymal and neural immunoreactivity.
        Development. 1990; 109: 461-471
        • Zhang J.
        • Zhang Y.
        • Lv H.
        • et al.
        Human stem cells from the apical papilla response to bacterial lipopolysaccharide exposure and anti-inflammatory effects of nuclear factor I C.
        J Endod. 2013; 39: 1416-1422
        • Vishwanat L.
        • Duong R.
        • Takimoto K.
        • et al.
        Effect of bacterial biofilm on the osteogenic differentiation of stem cells of apical papilla.
        J Endod. 2017; 43: 916-922
        • Ruparel N.B.
        • de Almeida J.F.
        • Henry M.A.
        • Diogenes A.
        Characterization of a stem cell of apical papilla cell line: effect of passage on cellular phenotype.
        J Endod. 2013; 39: 357-363
        • Livak K.J.
        • Schmittgen T.D.
        Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method.
        Methods. 2001; 25: 402-408
        • Pertea M.
        • Pertea G.M.
        • Antonescu C.M.
        • et al.
        StringTie enables improved reconstruction of a transcriptome from RNA-seq reads.
        Nat Biotechnol. 2015; 33: 290-295
        • Robinson M.D.
        • McCarthy D.J.
        • Smyth G.K.
        edgeR: a Bioconductor package for differential expression analysis of digital gene expression data.
        Bioinformatics. 2010; 26: 139-140
        • Ashburner M.
        • Ball C.A.
        • Blake J.A.
        • et al.
        Gene ontology: tool for the unification of biology. The Gene Ontology Consortium.
        Nat Genet. 2000; 25: 25-29
        • Kanehisa M.
        A database for post-genome analysis.
        Trends Genet. 1997; 13: 375-376
        • Luiz de Oliveira da Rosa W.
        • Machado da Silva T.
        • Fernando Demarco F.
        • et al.
        Could the application of bioactive molecules improve vital pulp therapy success? A systematic review.
        J Biomed Mater Res A. 2017; 105: 941-956
        • Levi-Montalcini R.
        The saga of the nerve growth factor.
        Neuroreport. 1998; 9: R71-R83
        • Yadlapati M.
        • Biguetti C.
        • Cavalla F.
        • et al.
        Characterization of a vascular endothelial growth factor-loaded bioresorbable delivery system for pulp regeneration.
        J Endod. 2017; 43: 77-83
        • Micera A.
        • Puxeddu I.
        • Lambiase A.
        • et al.
        The pro-fibrogenic effect of nerve growth factor on conjunctival fibroblasts is mediated by transforming growth factor-beta.
        Clin Exp Allergy. 2005; 35: 650-656
        • Nie X.
        • Tian W.
        • Zhang Y.
        • et al.
        Induction of transforming growth factor-beta 1 on dentine pulp cells in different culture patterns.
        Cell Biol Int. 2006; 30: 295-300
        • Takano S.
        • Uchida K.
        • Itakura M.
        • et al.
        Transforming growth factor-beta stimulates nerve growth factor production in osteoarthritic synovium.
        BMC Musculoskelet Disord. 2019; 20: 204
        • Kim S.J.
        • Park K.
        • Rudkin B.B.
        • et al.
        Nerve growth factor induces transcription of transforming growth factor-beta 1 through a specific promoter element in PC12 cells.
        J Biol Chem. 1994; 269: 3739-3744
        • Sone P.P.
        • Kaneko T.
        • Zaw S.Y.
        • et al.
        Neural regeneration/remodeling in engineered coronal pulp tissue in the rat molar.
        J Endod. 2020; 46: 943-949
        • Lei S.
        • Liu X.M.
        • Liu Y.
        • et al.
        Lipopolysaccharide downregulates the osteo-/odontogenic differentiation of stem cells from apical papilla by inducing autophagy.
        J Endod. 2020; 46: 502-508
        • Rosa V.
        • Zhang Z.
        • Grande R.H.
        • Nor J.E.
        Dental pulp tissue engineering in full-length human root canals.
        J Dent Res. 2013; 92: 970-975
        • Cardoso F.P.
        • Viana M.B.
        • Sobrinho A.P.
        • et al.
        Methylation pattern of the IFN-gamma gene in human dental pulp.
        J Endod. 2010; 36: 642-646
        • Campos K.
        • Franscisconi C.F.
        • Okehie V.
        • et al.
        FOXP3 DNA methylation levels as a potential biomarker in the development of periapical lesions.
        J Endod. 2015; 41: 212-218